CN113527383A - High-purity plant tannin prepared from industrial tannin extract and preparation method thereof - Google Patents
High-purity plant tannin prepared from industrial tannin extract and preparation method thereof Download PDFInfo
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Abstract
The invention discloses a high-purity plant tannin prepared by using industrial grade tannin extract and a preparation method thereof. The method comprises the following steps: (1) organic solvent treatment and degreasing of industrial grade tannin extract raw materials; (2) leaching the degreased industrial grade tannin extract residues by using an organic solvent and water to obtain a leaching solution; (3) concentrating the leaching liquor, extracting with an organic solvent to obtain an extract, and evaporating and drying to obtain a primarily refined plant tannin product with higher purity; (4) preparing the primarily refined solid product into a solution with a certain concentration, performing adsorption separation through macroporous resin subjected to swelling pretreatment, eluting with an eluant to obtain an eluent, and concentrating and drying to obtain a high-purity plant tannin product. The invention further improves the application value of the plant tannin product, widens the application field of the product, and has great social and economic significance for promoting the utilization of renewable resources and realizing the sustainable development of society.
Description
Technical Field
The invention belongs to the technical field of utilization of industrial tannin extracts, and relates to a high-purity plant tannin prepared from industrial tannin extracts and a preparation method thereof.
Background
Tannin extract (tannin extract) is a general name of a complex natural compound, and is mainly extracted from forestry wastes such as skin, root, leaf, shell and the like of plant bodies, wherein non-tannin and insoluble substances are contained in the tannin besides main components of plant bodies. The compositions are different due to different raw materials. The trade name is generally preceded by the name of raw materials, such as larch bark extract, valonia extract, myrica rubra extract and the like, so as to distinguish the composition, the property and the application.
The specific preparation process of the industrial-grade myrica rubra tannin extract comprises the following steps: crushing, screening and removing impurities to obtain crushed materials (the granularity is 2.0-20mm, usually 1.5-3.0mm), leaching the crushed materials with hot water (the temperature is 90-120 ℃, the using amount of sodium sulfite is 1.0-1.8% (accounting for air-dried raw materials), the number of tanks is 6-10, the time is 12-20h, leaching is 15-19 times, the liquid outlet coefficient is 600 plus 650 percent) to obtain leaching liquor, filtering and clarifying the leaching liquor to obtain purified leaching liquor, evaporating and concentrating the purified leaching liquor (multi-effect evaporation, controlling the concentration of concentrated glue liquor to be 30-55 percent) to obtain concentrated glue liquor, and spray drying the concentrated glue liquor to obtain the reddish brown group powder. The first-class product contains tannin with the mass fraction not less than 67%.
Plant tannin is also called plant polyphenol, is a complex phenol secondary metabolite in a plant body, has a polyphenol structure, and is rich in the reserve of plant polyphenol in nature. Tannin can be classified into soluble tannin and insoluble tannin due to their different solubilities, and the main substance considered to cause astringency is soluble tannin, which produces strong astringency in combination with oral mucosa proteins of the human body, thereby causing "astringency". According to the chemical structure characteristics of tannin, the main components of tannin extract can be divided into:
first, tannin-like substances are hydrolyzed. Complex compounds of polyphenol carboxylic acids bound to sugars, such as glucose or polyols, by ester or glycoside bonds can be hydrolyzed by acid or enzyme to the corresponding products. According to the structure of polyphenol carboxylic acid obtained after hydrolysis, the polyphenol carboxylic acid is divided into gallotannin (or gall tannin contained in gallnut and the like) and ellagitannin (tannin contained in valonia and the like), and the structural formula is shown in figure 1 (Guolinxin and the like, the research on the structural modification of plant tannin, chemical science and report, https:// knns, cnki, net/kcms/detail/11.1946, TQ.20201218.1000.004.html). The hydrolyzed plant tannin is widely applied to the fields of food, medical drugs, beverages, cosmetics and the like, and has the most obvious effect in the medical industry.
② condensed tannins, also called condensed tannins, which are complex condensates formed by the main combination of compounds such as flavanols and the like with carbon-carbon bonds. In aqueous solution, it cannot be hydrolyzed by acid or enzyme; in contrast, the co-heating with acid condenses into an amorphous material that is poorly soluble in water, red powder. Most monomers in condensed tannin molecules are derived from flavonoid compounds such as hydroxy flavan-3-ol, flavan-3, 4-diol and the like, and the structural formula is shown in figure 2 (Guolinxin and the like. structural modification research of plant tannin, chemical science report. https:// kns.cnki.net/kcms/detail/11.1946.TQ.20201218.1000.004. html). Therefore, condensed tannin is also called flavonoid tannin, and is a part of the original xanthic pigment in the flavonoid compounds. In nature, condensed tannins are much more abundant than hydrolyzed tannins, and in the yield of tannins in the world, condensed tannins account for over 90%, but the practical application of condensed tannins is less extensive than that of hydrolyzed tannins.
The tannin extract contains main component tannin, and also contains partial non-tannin components and insoluble substances, wherein the non-tannin components comprise sugar, simple phenol, organic acid, inorganic salt, pigment, nitrogen-containing substances, and the like. The non-tannin compositions of the various tannin extracts are different. The insoluble substances mainly include tannin decomposition product or condensation product, pectin, gum and low-dispersion tannin, impurities and inorganic salt. The vegetable tannin with higher purity is difficult to obtain from the tannin extract because the tannin extract has more soluble components, small molecular weight difference and difficult separation and purification.
The tannin extract also contains a small amount of insoluble substances, i.e. substances which can not pass through the medium-speed filter paper and the kaolin filter layer when the tannin solution with the concentration of 0.4% is at the temperature of 20 +/-2 ℃. The main components are decomposition product (yellow powder) or condensation product (red powder) of tannin, low-dispersity tannin, pectin, gum, inorganic salt, mechanical impurities, etc.
According to the industry standard LY/T1084-.
TABLE 1 technical index of Chinese waxberry tannin extract
| Index name | Superior product | First-class product | Qualified product |
| Tannin mass fraction/% > or more | 69 | 67 | 65 |
| Insoluble matter/% < or less | 4 | 5 | 6 |
| Moisture content is less than or equal to | 12 | 12 | 12 |
| Precipitation/% < or | 2 | 4 | 6 |
| pH value | 4.5-5.5 | 4.5-5.5 | 4.5-5.5 |
The plant tannin in China is not only very rich in resources, but also very rich in varieties, for example, the plants such as mangrove, acacia mangosteen, black wattle, myrica esculenta, stone plants, ephedra, rhubarb, gallnut, larch, tea tree and the like which are wide in planting distribution area and high in economic value are all natural resources rich in the plant tannin.
The plant tannin can be combined with protein through hydrophobic bond and multi-point hydrogen bond to react, which is the most important chemical characteristic. Similar to the molecular complex reaction of plant polyphenol and other biological macromolecules, such as alkaloid, polysaccharide, etc. The complex reaction between several ortho-phenolic hydroxyl radicals in plant polyphenol and metal ion is the chemical basis for several applications. Antioxidant properties are also an important property of plant polyphenols. Because the ortho-position phenolic hydroxyl in the phenolic hydroxyl of the plant polyphenol is easy to oxidize and has stronger capturing capability on free radicals such as active oxygen, the plant polyphenol has strong oxidation resistance and free radical scavenging capability. In addition, the plant polyphenol has stronger ultraviolet light absorption capacity between 200-300 nm. With the continuous and deep research work of the structure and the chemical property of the plant polyphenol, people gradually deepen the understanding of the plant polyphenol and simultaneously lay a theoretical foundation for the application of the plant polyphenol in various fields.
Hydrolyzed tannins and condensed tannins are completely different in structural composition, and thus they are significantly different in chemical properties and application range. For example, the hydrolyzed tannin is unstable under the action of acid, alkali and enzyme and is easy to hydrolyze; condensed tannins are relatively stable, but condense into water-insoluble substances under the action of strong acid. The hydrolyzed tannins and condensed tannins still have some commonalities in molecular structure, such as the two contain abundant phenolic hydroxyl groups, and are most typical of ortho-phenolic hydroxyl groups; the molecular weight is larger and the distribution is wider. It is this chemical structure that imparts unique chemical properties to polyphenols. The tannin content in tannin extract is measured by detecting the total phenol content.
At present, no method for obtaining high-purity plant tannin from industrial grade tannin extract is reported.
Aiming at the research of the content of the main component plant tannin and other component compositions and properties in the industrial tannin extract, the invention adopts the following steps: (1) degreasing with an organic solvent; (2) further leaching the residual filter residue of the degreased industrial grade tannin extract by using an organic solvent and water to obtain a leaching solution; (3) concentrating the leaching liquor, extracting with an organic solvent to obtain an extract, separating and drying to obtain a preliminarily refined plant tannin solid product with higher purity; (4) preparing a plant tannin solid product with higher purity obtained by primary refining into a solution with a certain concentration, and further performing adsorption separation by using macroporous resin (subjected to swelling treatment in advance); (5) washing the chromatographic column with distilled water, eluting with ethanol solution, and collecting the eluate; (6) concentrating and drying the eluent to obtain a red-black solid, namely obtaining the plant tannin product with high purity. Further improves the application value of the product, widens the application field of the product, and has great social and economic significance for promoting the utilization of renewable resources and realizing the sustainable development of society.
Disclosure of Invention
The invention aims to provide a method for preparing a high-purity plant tannin product by separating and purifying an industrial grade tannin extract, which is suitable for the application field of products needing plant tannin with higher purity or high purity as raw materials or components, can reduce the influence of impurity components and improve the service performance.
In order to solve the technical problem, the invention provides a method for purifying an industrial-grade tannin extract, which comprises the following steps:
(1) adding petroleum ether into industrial tannin extract, mixing well, degreasing under ultrasonic condition, separating, and collecting filtrate and residue respectively;
(2) drying the filter residue obtained in the step (1), adding acetone and distilled water into the dried filter residue, fully stirring, separating, and collecting filtrate;
(3) removing acetone from the filtrate collected in the step (2), collecting concentrated solution, extracting with n-hexane, collecting lower layer liquid, extracting with chloroform, collecting upper layer liquid, and drying to obtain primarily refined plant tannin solid;
(4) adding water into the primarily refined plant tannin solid obtained in the step (3) to prepare a primarily refined plant tannin solution;
(5) passing the primarily refined plant tannin solution prepared in the step (4) through a pretreated macroporous adsorption resin chromatographic column;
(6) after the adsorption balance in the step (5) is achieved, firstly, washing the chromatographic column with water, then, eluting with ethanol solution as an eluent, and collecting the eluent;
(7) and (4) concentrating the eluent collected in the step (6), and drying to obtain the plant tannin product.
The method comprises the following steps: the industrial grade tannin extract (first-class product) in the step (1) is prepared by taking myrica rubra barks as raw materials. The method specifically comprises the steps of crushing, hot water leaching, filtering and spray drying to obtain reddish brown group powder, wherein the mass fraction of tannin is not less than 67%.
The specific preparation process of the industrial-grade myrica rubra tannin extract comprises the following steps: crushing, screening and removing impurities to obtain crushed materials (the granularity is 2.0-20mm, usually 1.5-3.0mm), leaching the crushed materials with hot water (the temperature is 90-120 ℃, the using amount of sodium sulfite is 1.0-1.8% (accounting for air-dried raw materials), the number of tanks is 6-10, the time is 12-20h, leaching is 15-19 times, the liquid outlet coefficient is 600 plus 650 percent) to obtain leaching liquor, filtering and clarifying the leaching liquor to obtain purified leaching liquor, evaporating and concentrating the purified leaching liquor (multi-effect evaporation, controlling the concentration of concentrated glue liquor to be 30-55 percent) to obtain concentrated glue liquor, and spray drying the concentrated glue liquor to obtain the reddish brown group powder. The first-class product contains tannin with the mass fraction not less than 67%.
The method comprises the following steps: the boiling range of the petroleum ether used in the step (1) is 60-90 ℃, and the solid-to-liquid ratio of the industrial-grade tannin extract to the petroleum ether is 1:1-1:4(g/mL), and the preferred solid-to-liquid ratio is 1:2 (g/mL).
The method comprises the following steps: the degreasing time in the step (1) is 0.5-1.0h, and then filtration or centrifugal separation is carried out to respectively obtain filtrate and filter residue; the ultrasonic power range 240-.
The method comprises the following steps: and (2) drying the filter residue obtained in the step (1) until the water content is less than or equal to 2%, wherein the volume ratio of acetone to distilled water is 1:9-9:1, the total solid-liquid ratio is 1:5-1:15(g: mL), preferably the volume ratio of acetone to distilled water is 7:3, the total solid-liquid ratio is 1:10(g: mL), fully stirring for 0.5-1.0h, filtering or centrifugally separating, and collecting the filtrate.
The method comprises the following steps: removing acetone from the filtrate obtained in the step (2) by using a rotary evaporator at the vacuum degree of 95-98 kPa and the heating bath temperature of 38-42 ℃, and collecting a concentrated solution with the mass concentration controlled at 28-32%; and then extracting for at least 1 time by using n-hexane with the same volume, collecting lower layer liquid, extracting for at least 1 time by using chloroform with the same volume, collecting upper layer liquid, and then carrying out vacuum drying at 30-60 ℃, preferably 40 ℃ until the water content is less than or equal to 2%, thus obtaining red-black solid, namely the primarily refined plant tannin solid. The product yield can reach more than 60 percent, and the total phenol content can reach more than 83 percent.
The method comprises the following steps: in the step (4), the preliminarily refined plant tannin solid sample obtained in the step (3) is added with water to prepare a solution with the mass concentration of 1.0g/L-5.0g/L, preferably 1.5g/L-3.0 g/L.
The method comprises the following steps: step (5) putting the pretreated macroporous resin into a chromatographic column by a wet method, adding the preliminarily refined plant tannin solution prepared in the step (4) of 1.0BV-6.0BV, and adsorbing at the flow rate of 1.0BV/h-4.0 BV/h; preferably, 4.0BV of the primarily refined plant tannin solution prepared in the step (4) is added, and the adsorption is carried out at the flow rate of 1.5 BV/h;
after the adsorption balance is achieved in the step (6), washing the chromatographic column by distilled water with the volume of 1.0BV-4.0BV, then eluting by using ethanol solution with the volume fraction of 1.0BV-3.0BV being 40% -90% as an eluent with the flow rate being 1.0-3.0BV/h, and collecting eluent; preferably 2.0BV volume of distilled water is used to wash the column, preferably 1.6BV volume fraction 70% ethanol solution is used as eluent, and the flow rate is preferably 1.5 BV/h.
The method comprises the following steps: and (7) concentrating the eluent obtained in the step (6) at 30-60 ℃ by using a vacuum drying device, preferably at 50 ℃, and then drying at 30-60 ℃, preferably at 40 ℃ until the water content is less than or equal to 2% to obtain a plant tannin solid sample.
The invention also provides the plant tannin prepared by the method.
The high-purity plant tannin product obtained after further refining and purification has the total product yield of more than 20 percent and the total phenol content of more than 96 percent (determined by a Prussian blue method).
The macroporous resin (such as HPD100 type and HPD826 type macroporous adsorption resin) pretreatment process adopted in the method comprises the following steps: soaking in ethanol solution to swell the macroporous adsorbent resin, filtering, washing with distilled water until the filtrate has no alcohol smell, and soaking in distilled water.
The invention prepares the high-purity plant tannin product by utilizing the combined separation and purification of an industrial grade tannin extract raw material by an organic solvent method and a macroporous resin adsorption method, solves the problem that the industrial tannin extract product has common use performance or is difficult to be applied to the field with higher requirement on the purity of the plant tannin due to lower content of effective components of the plant tannin, enlarges the use range of the product and improves the added value of the product.
Drawings
FIG. 1 shows the composition and structure of hydrolyzed tannin;
FIG. 2 shows the composition and structure of condensed tannin;
FIG. 3 is a graph showing a standard curve for measuring the total phenol content;
the abscissa is the concentration of the tannic acid solution (mg/mL) and the ordinate is the absorbance of the ultraviolet spectrum.
Detailed Description
The present invention is further illustrated by the following examples, which are not to be construed as limiting the invention.
Industrial myrica rubra tannin extract and determination of total phenol content of each purified sample
According to the literature, the total phenol content of each sample, namely the content of tannin in a representative sample, is determined by drawing a standard curve for measuring the total phenol concentration by using the prussian blue method and using tannic acid as a standard sample at 700nm by using an ultraviolet spectrophotometer, and measuring the absorbance of the myrica rubra extract and the samples obtained by each purification method by using the same conditions and methods. The method comprises the following specific steps:
(1) weighing 0.1000g of tannic acid standard sample, adding a proper amount of distilled water for dissolving, and fixing the volume to a 100mL volumetric flask to obtain the tannic acid solution A with the concentration of 1.00 mg/mL. Then measuring 1.00mL, 2.00mL, 3.00mL, 4.00mL and 5.00mL of tannic acid standard solution A by a pipette respectively, and metering the volume to a 100mL volumetric flask to obtain 5 gradient tannic acid standard solutions B with the volume of 0.01 mg/mL-0.05 mg/mL.
Taking a tannic acid standard sample of 0.01mg/mL as an example, the specific flow of the Prussian blue method is as follows: accurately transferring 1.00mL of tannic acid standard solution with the mass concentration of 0.01mg/mL, adding 1.00mL of potassium ferricyanide solution with the molar concentration of 0.016mol/L, fully mixing uniformly, placing at (24 +/-1) DEG C for reaction for 15min, immediately adding 1.00mL of 0.02mol/L ferric trichloride solution iron solution prepared by 1.00mol/L hydrochloric acid, subsequently adding 3.00mL of phosphoric acid solution with the molar concentration of 6.02mol/L, fully mixing uniformly to terminate the reaction, dropwise adding 2.00mL of Arabic gum with the mass concentration of 1%, fully mixing uniformly to prevent the generation of precipitates, and measuring the absorbance of a sample at 700nm by using an ultraviolet spectrophotometer.
A blank sample was prepared from deionized water according to the above method, and the absorbance of tannic acid standard sample B at each concentration was measured, thereby drawing a standard curve for the measurement of total phenol concentration, as shown in FIG. 3. Regression linear equation: y is 9.96X +0.2786 and R is 0.9998.
(2) Determination of total phenol content of purified tannin samples prepared by each separation and purification method: weighing 0.2000g of plant tannin sample or industrial myrica rubra tannin extract raw material which is treated by various purification methods, preparing sample solution, transferring 10.00mL of the solution into a surface dish, and drying at 40 ℃ to obtain the solid content of 10.00mL of the solution; then, 2.00mL of each sample solution was removed and the volume was determined in a 100mL volumetric flask. And (3) measuring the absorbance of each sample solution under the same experimental method and conditions as those for preparing the total phenol concentration measurement standard curve, and calculating the total phenol content of each sample according to the standard curve, namely representing the content of the plant tannin in each sample.
I, calculation formula of total phenol content of sample
Samples treated by various purification methods were weighed and the total phenol concentration was recordedC1(mg/mL) of the solution, 10.00mL of the solution was transferred to a petri dish, the total phenol mass was denoted as N (mg), and the solution was dried at 70-80 ℃ to obtain a solid mass of 10.00mL of the solution, denoted as M (mg); then 2.00mL of the solution is transferred into a 100mL volumetric flask for dilution and constant volume, and the total phenol concentration of the diluted solution is marked as C2(mg/mL). Measuring the absorbance A of each sample solution under the same experimental method and conditions as the total phenol concentration measurement standard curve prepared above, and calculating to obtain the total phenol concentration C of each sample in each dilution according to the standard curve2(mg/mL). Finally, the relative content of total phenols in each sample is calculated as follows:
C1(mg/mL)＝C2(mg/mL)×100(mL)÷2(mL)
N(mg)＝C1(mg/mL)×10(mL)
the total phenol content of each sample was N (mg)/M (mg) × 100%
II, calculation formula of sample yield
Sample yield (%) - [ product mass (g)/raw material mass (g) obtained after separation and purification ]. times.100%
Comparative example 1
Taking 10g of industrial myrica rubra tannin extract (first-grade product), adding 80% (V/V) of anhydrous ethanol and ethyl acetate aqueous solution (wherein the volume ratio of ethanol to ethyl acetate is 1:1) according to the solid-to-liquid ratio of 1:10(g/ml), carrying out ultrasonic treatment for 30min at 50 ℃ and 350w, then carrying out suction filtration, collecting filtrate, concentrating at 40 ℃, carrying out forced air drying at 40 ℃ until the water content is less than or equal to 2%, wherein the yield of the obtained product is 51.30%, and the total phenol content is 56.91%.
Comparative example 2
Taking 10g of industrial myrica rubra tannin extract (first-grade product), adding 80% (V/V) of anhydrous ethanol and ethyl acetate aqueous solution (wherein the volume ratio of ethanol to ethyl acetate is 1:1) according to the solid-to-liquid ratio of 1:6(g/ml), carrying out ultrasonic treatment for 30min at 50 ℃ and 350w, then carrying out suction filtration, collecting filtrate, concentrating at 40 ℃, carrying out blast drying at 40 ℃ until the water content is less than or equal to 2%, wherein the yield of the obtained product is 53.56%, and the total phenol content is 52.72%.
Comparative example 3
Adding petroleum ether into 10g of industrial myrica rubra tannin extract (first-grade product) according to the mass concentration ratio of 1:3(g/mL), fully shaking up, degreasing for 30min under 350w ultrasound, filtering to remove the petroleum ether, and drying by air blast at 40 ℃ (the water content is less than or equal to 2%) to obtain the degreased myrica rubra tannin extract. Adding 60% ethanol solution (V/V) into 5g of cortex myricae Precatori extract at a solid-to-liquid ratio of 1:10(g/ml), stirring thoroughly for dissolving, filtering, and air drying the solid residue at 40 deg.C (water content is less than or equal to 2%), wherein the product yield is 58.34%, and the total phenol content of the residue is 52.72%.
Comparative example 4
Taking 10g of industrial myrica rubra tannin extract (first-grade product), adding water and acetone (the volume ratio of water to acetone is 3:7) according to the solid-to-liquid ratio of 1:5(g/mL), dissolving, fully stirring, filtering and collecting filtrate, removing the acetone in the filtrate by using a rotary evaporator at the vacuum degree of 95-98 kPa and the heating bath temperature of about 40 ℃, collecting the filtrate after removing the acetone, and drying the filtrate in a vacuum drying oven at the temperature of 40-50 ℃ to obtain a red-black solid sample, wherein the yield of the product is 64.88%, and the total phenol content of the product is 44.54%.
Comparative example 5
Taking 10g of industrial myrica rubra tannin extract (first-grade product), adding water and acetone (the volume ratio of water to acetone is 3:7) according to the solid-to-liquid ratio of 1:7.5g/mL, dissolving, fully stirring, filtering and collecting filtrate, removing the acetone in the filtrate by using a rotary evaporator at the vacuum degree of 95-98 kPa and the heating bath temperature of about 40 ℃, collecting concentrated solution (the mass concentration is controlled to be about 30%) from which the acetone is removed, and drying the concentrated solution in a vacuum drying oven at the temperature of 40-50 ℃ (the water content is less than or equal to 2%) to obtain a red-black solid sample, wherein the yield of the product is 59.17%, and the total phenol content of the product is 71.15%.
Comparative example 6
Adding petroleum ether into 10g of industrial myrica rubra tannin extract (first-grade product) according to the mass-volume ratio of 1:2(g/mL), fully shaking, degreasing for 30min under ultrasonic, filtering to remove petroleum ether, and drying with blast at 40 ℃ (water content is less than or equal to 2%). Adding 70.00mL of acetone into the degreased myrica rubra tannin extract, slowly adding 30.00mL of distilled water, fully stirring for 30min, filtering by using a triangular funnel, and collecting the filtrate. Removing acetone from the filtrate by using a rotary evaporator at the vacuum degree of 95-98 kPa and the heating bath temperature of about 40 ℃, collecting a concentrated solution (the mass concentration is controlled to be about 30%), extracting for 1 time by using n-hexane with the same volume, collecting the lower raffinate, extracting for 1 time by using chloroform with the same volume, and collecting the upper liquid. Drying at 40 deg.C (water content is less than or equal to 2%) to obtain primary refined tannin product with yield of 60.13% and total phenol content of 83.10%.
Comparative example 7
Adding petroleum ether into 10g of industrial myrica rubra tannin extract (first-grade product) according to the mass-volume ratio of 1:2(g/mL), fully shaking, degreasing for 30min under ultrasonic, filtering to remove petroleum ether, and drying with blast at 40 ℃ (water content is less than or equal to 2%). Adding 70.00mL of acetone into degreased myrica rubra tannin extract, slowly adding 30.00mL of distilled water, fully stirring for 30min, filtering by using a triangular funnel, and collecting filtrate. Removing acetone from the filtrate by using a rotary evaporator at the vacuum degree of 95-98 kPa and the heating bath temperature of about 40 ℃, collecting a concentrated solution (the mass concentration is controlled to be about 30%), extracting for 2 times by using n-hexane with the same volume, collecting the lower raffinate, extracting for 2 times by using chloroform with the same volume, and collecting the upper liquid. Drying at 40 deg.C (water content is less than or equal to 2%) to obtain refined tannin product with yield of 52.54% and total phenol content of 91.42%.
Comparative example 8
Adding petroleum ether into 10g of industrial myrica rubra tannin extract (first-grade product) according to the mass-volume ratio of 1:2(g/mL), fully shaking, degreasing for 30min under ultrasonic, filtering to remove petroleum ether, and drying with blast at 40 ℃ (water content is less than or equal to 2%). Adding 70.00mL of acetone into degreased myrica rubra tannin extract, slowly adding 30.00mL of distilled water, fully stirring for 30min, filtering by using a triangular funnel, and collecting filtrate. Removing acetone from the filtrate by using a rotary evaporator at the vacuum degree of 95-98 kPa and the heating bath temperature of about 40 ℃, collecting a concentrated solution (the mass concentration is controlled to be about 30%), extracting for 3 times by using n-hexane with the same volume, collecting the lower raffinate, extracting for 3 times by using chloroform with the same volume, and collecting the upper liquid. Drying at 40 deg.C (water content is less than or equal to 2%) to obtain primary refined tannin product with yield of 50.35% and total phenol content of 93.03%.
Comparative example 9
30g of HPD100 type macroporous adsorption resin is taken, 95% ethanol is added for soaking for 24h to ensure that the macroporous adsorption resin is fully swelled, the filtration is carried out, then a large amount of distilled water is used for washing until the filtered solution has no obvious alcohol smell, and the distilled water is used for soaking and storing for later use.
30g of the treated HPD100 type resin is taken, wet loading is carried out on the resin, and adsorption is carried out by adding 4.0BV1.20g/L of industrial myrica rubra extract solution (same as the raw material, the first-class product and the same below) at the flow rate of 1.5 BV/h. After the adsorption equilibrium, the column was washed with 2.0BV of distilled water and eluted with 1.6BV of 70% ethanol solution at a flow rate of 1.5 BV/h. Collecting eluate, concentrating at 50 deg.C with rotary evaporator, and drying at 40 deg.C (water content is less than or equal to 2%) to obtain primarily refined tannin product with yield of 45.00% and total phenol content of 86.16%.
Comparative example 10
Taking 30g of HPD826 type macroporous adsorption resin, adding 95% ethanol, soaking for 24h to fully swell the macroporous adsorption resin, filtering, washing with a large amount of distilled water until the filtered solution has no obvious alcohol smell, and soaking with distilled water for storage.
30g of the treated HPD826 type resin is taken and loaded into a chromatographic column by a wet method, and 4.0BV1.20g/L of industrial myrica rubra extract solution is added for adsorption at the flow rate of 1.5 BV/h. After the adsorption equilibrium, the column was washed with 2.0BV of distilled water and eluted with 1.6BV of 70% ethanol solution at a flow rate of 1.5 BV/h. Collecting eluate, concentrating at 50 deg.C with rotary evaporator, and drying at 40 deg.C (water content is less than or equal to 2%) to obtain primarily refined tannin product with yield of 46.67% and total phenol content of 91.66%.
Comparative example 11
30g of HPD100 type macroporous adsorption resin is taken, 95% ethanol is added for soaking for 24h to ensure that the macroporous adsorption resin is fully swelled, the filtration is carried out, then a large amount of distilled water is used for washing until the filtered solution has no obvious alcohol smell, and the distilled water is used for soaking and storing for later use.
30g of the treated HPD100 type resin is taken, wet loading is carried out on the resin in a chromatographic column, and 4.0BV2.00g/L of industrial myrica rubra extract solution is added for adsorption at the flow rate of 1.5 BV/h. After the adsorption equilibrium, the column was washed with 2.0BV of distilled water and eluted with 1.6BV of 70% ethanol solution at a flow rate of 1.5 BV/h. Collecting eluate, concentrating at 50 deg.C with rotary evaporator, and drying at 40 deg.C (water content is less than or equal to 2%) to obtain primarily refined tannin product with yield of 43.00% and total phenol content of 84.54%.
Comparative example 12
Taking 30g of HPD826 type macroporous adsorption resin, adding 95% ethanol, soaking for 24h to fully swell the macroporous adsorption resin, filtering, washing with a large amount of distilled water until the filtered solution has no obvious alcohol smell, and soaking with distilled water for storage.
30g of the treated HPD826 type resin is taken and loaded into a chromatographic column by a wet method, and 4.0BV2.00g/L of industrial myrica rubra extract solution is added for adsorption at the flow rate of 1.5 BV/h. After the adsorption equilibrium, the column was washed with 2.0BV of distilled water and eluted with 1.6BV of 70% ethanol solution at a flow rate of 1.5 BV/h. Collecting eluate, concentrating at 50 deg.C with rotary evaporator, and drying at 40 deg.C (water content is less than or equal to 2%) to obtain primarily refined tannin product with yield of 46.00% and total phenol content of 85.32%.
Comparative example 13
30g of HPD100 type macroporous adsorption resin is taken, 95% ethanol is added for soaking for 24h to ensure that the macroporous adsorption resin is fully swelled, the filtration is carried out, then a large amount of distilled water is used for washing until the filtered solution has no obvious alcohol smell, and the distilled water is used for soaking and storing for later use.
30g of the treated HPD100 type resin is taken, wet loading is carried out on the resin in a chromatographic column, and 4.0BV3.00g/L of industrial myrica rubra extract solution is added for adsorption at the flow rate of 1.5 BV/h. After the adsorption equilibrium, the column was washed with 2.0BV of distilled water and eluted with 1.6BV of 70% ethanol solution at a flow rate of 1.5 BV/h. Collecting eluate, concentrating at 50 deg.C with rotary evaporator, and drying at 40 deg.C (water content is less than or equal to 2%) to obtain primarily refined tannin product with yield of 46.67% and total phenol content of 82.68%.
Comparative example 14
Taking 30g of HPD826 type macroporous adsorption resin, adding 95% ethanol, soaking for 24h to fully swell the macroporous adsorption resin, filtering, washing with a large amount of distilled water until the filtered solution has no obvious alcohol smell, and soaking with distilled water for storage.
30g of the treated HPD826 type resin is taken and loaded into a chromatographic column by a wet method, and 4.0BV3.00g/L of industrial myrica rubra extract solution is added for adsorption at the flow rate of 1.5 BV/h. After the adsorption equilibrium, the column was washed with 2.0BV of distilled water and eluted with 1.6BV of 70% ethanol solution at a flow rate of 1.5 BV/h. Collecting eluate, concentrating at 50 deg.C with rotary evaporator, and drying at 40 deg.C (water content is less than or equal to 2%) to obtain primarily refined tannin product with yield of 50.57% and total phenol content of 83.48%.
Comparative example 15
Taking 30g of HPD826 type macroporous adsorption resin, adding 95% ethanol, soaking for 24h to fully swell the macroporous adsorption resin, filtering, washing with a large amount of distilled water until the filtered solution has no obvious alcohol smell, and soaking with distilled water for storage.
30g of the treated HPD826 type resin is taken and loaded into a chromatographic column by a wet method, and 6.0BV3.00g/L of industrial myrica rubra extract solution is added for adsorption at the flow rate of 1.5 BV/h. After the adsorption equilibrium, the column was washed with 2.0BV of distilled water and eluted with 1.6BV of 70% ethanol solution at a flow rate of 1.5 BV/h. Collecting eluate, concentrating at 50 deg.C with rotary evaporator, and drying at 40 deg.C (water content is less than or equal to 2%) to obtain primarily refined tannin product with yield of 47.23% and total phenol content of 81.75%.
Comparative example 16
Weighing 30g of HPD826 type macroporous adsorption resin, adding 95% ethanol, soaking for 24h to fully swell the macroporous adsorption resin, filtering, washing with a large amount of distilled water until the filtered solution has no obvious alcohol smell, and soaking with distilled water for storage.
30g of the treated HPD826 type resin is taken and loaded into a chromatographic column by a wet method, and 2.0BV3.00g/L of industrial myrica rubra extract solution is added for adsorption at the flow rate of 1.5 BV/h. After the adsorption equilibrium, the column was washed with 2.0BV of distilled water and eluted with 1.6BV of 70% ethanol solution at a flow rate of 1.5 BV/h. Collecting eluate, concentrating at 50 deg.C with rotary evaporator, and drying at 40 deg.C (water content is less than or equal to 2%) to obtain primarily refined tannin product with yield of 54.23% and total phenol content of 87.32%.
Comparative example 17
Taking 30g of HPD826 type macroporous adsorption resin, adding 95% ethanol, soaking for 24h to fully swell the macroporous adsorption resin, filtering, washing with a large amount of distilled water until the filtered solution has no obvious alcohol smell, and soaking with distilled water for storage.
30g of the treated HPD826 type resin is taken and loaded into a chromatographic column by a wet method, and 4.0BV2.00g/L of industrial myrica rubra extract solution is added for adsorption at the flow rate of 1.5 BV/h. After the adsorption equilibrium, the column was washed with 2.0BV of distilled water and eluted with 3.0BV of 70% ethanol at a flow rate of 1.5 BV/h. Collecting eluate, concentrating at 50 deg.C with rotary evaporator, and drying at 40 deg.C (water content is less than or equal to 2%) to obtain primarily refined tannin product with yield of 49.56% and total phenol content of 87.53%.
Comparative example 18
Taking 30g of HPD826 type macroporous adsorption resin, adding 95% ethanol, soaking for 24h to fully swell the macroporous adsorption resin, filtering, washing with a large amount of distilled water until the filtered solution has no obvious alcohol smell, and soaking with distilled water for storage.
30g of the treated HPD826 type resin is taken and loaded into a chromatographic column by a wet method, and 4.0BV2.00g/L of industrial myrica rubra extract solution is added for adsorption at the flow rate of 1.5 BV/h. After the adsorption equilibrium, the column was washed with 2.0BV of distilled water and eluted with 1.0BV of 70% ethanol solution at a flow rate of 1.5 BV/h. Collecting eluate, concentrating at 50 deg.C with rotary evaporator, and drying at 40 deg.C (water content is less than or equal to 2%) to obtain primarily refined tannin product with yield of 44.72% and total phenol content of 88.14%.
Comparative example 19
Taking 30g of HPD826 type macroporous adsorption resin, adding 95% ethanol, soaking for 24h to fully swell the macroporous adsorption resin, filtering, washing with a large amount of distilled water until the filtered solution has no obvious alcohol smell, and soaking with distilled water for storage.
30g of the treated HPD826 type resin is taken and loaded into a chromatographic column by a wet method, and 4.0BV2.00g/L of industrial myrica rubra extract solution is added for adsorption at the flow rate of 1.5 BV/h. After the adsorption equilibrium, the column was washed with 2.0BV of distilled water and eluted with 1.6BV of 70% ethanol solution at a flow rate of 1.0 BV/h. Collecting eluate, concentrating at 50 deg.C with rotary evaporator, and drying at 40 deg.C (water content is less than or equal to 2%) to obtain primarily refined tannin product with yield of 48.53% and total phenol content of 86.35%.
Comparative example 20
Taking 30g of HPD826 type macroporous adsorption resin, adding 95% ethanol, soaking for 24h to fully swell the macroporous adsorption resin, filtering, washing with a large amount of distilled water until the filtered solution has no obvious alcohol smell, and soaking with distilled water for storage.
30g of the treated HPD826 type resin is taken and loaded into a chromatographic column by a wet method, and 4.0BV2.00g/L of industrial myrica rubra extract solution is added for adsorption at the flow rate of 1.5 BV/h. After the adsorption equilibrium, the column was washed with 2.0BV of distilled water and eluted with 1.6BV of 70% ethanol at a flow rate of 3.0 BV/h. Collecting eluate, concentrating at 50 deg.C with rotary evaporator, and drying at 40 deg.C (water content is less than or equal to 2%) to obtain primarily refined tannin product with yield of 45.14% and total phenol content of 84.77%.
Comparative example 21
Adding petroleum ether into 10g of industrial myrica rubra tannin extract according to the mass-volume ratio of 1:2(g/mL), fully shaking up, degreasing for 30min under ultrasonic, filtering to remove the petroleum ether, and drying the myrica rubra tannin extract by blast at 40 ℃ (the water content is less than or equal to 2%). Adding 70mL of acetone into the degreased myrica rubra tannin extract, then slowly adding 30mL of distilled water, fully stirring for 30min, filtering by using a triangular funnel and collecting the filtrate. Removing acetone from the filtrate by using a rotary evaporator at the vacuum degree of 95-98 kPa and the heating bath temperature of about 40 ℃, collecting a concentrated solution (the mass concentration is controlled to be about 30%), extracting for 1 time by using n-hexane with the same volume, collecting the lower raffinate, extracting for 1 time by using chloroform with the same volume, and collecting the upper liquid. Drying at 40 deg.C (water content is less than or equal to 2%) to obtain primary refined tannin product with yield of 60.13% and total phenol content of 83.10%.
Taking 30g of HPD826 type macroporous adsorption resin, adding 95% ethanol, soaking for 24h to fully swell the macroporous adsorption resin, filtering, washing with a large amount of distilled water until the filtered solution has no obvious alcohol smell, and soaking with distilled water for storage.
30g of the treated HPD826 type resin is taken and put into a chromatographic column by a wet method, and 8.0BV of tannin solution of 3.00g/L prepared by the above primary refined tannin sample is added for adsorption at a flow rate of 1.5 BV/h. After the adsorption equilibrium, the column was washed with 2.0BV of distilled water and eluted with 1.6BV of 70% ethanol solution at a flow rate of 1.5 BV/h. Collecting eluate, concentrating at 50 deg.C with rotary evaporator, and drying at 40 deg.C (water content is less than or equal to 2%) to obtain refined tannin product with yield of 44.24%, total yield of refined tannin product of 26.60%, and total phenol content of refined tannin product of 92.62%.
Example 1
Taking 10g of industrial myrica rubra tannin extract (first-class product, same as above and same as below), adding petroleum ether according to the mass-volume ratio of 1:2(g/mL), fully shaking up, degreasing for 30min under ultrasonic, filtering to remove the petroleum ether, and drying the myrica rubra tannin extract by blast at 40 ℃ (the water content is less than or equal to 2%). Adding 70mL of acetone into the degreased myrica rubra tannin extract, then slowly adding 30mL of distilled water, fully stirring for 30min, filtering by using a triangular funnel and collecting the filtrate. Removing acetone from the filtrate by using a rotary evaporator at the vacuum degree of 95-98 kPa and the heating bath temperature of about 40 ℃, collecting a concentrated solution (the mass concentration is controlled to be about 30%), extracting for 1 time by using n-hexane with the same volume, collecting the lower raffinate, extracting for 1 time by using chloroform with the same volume, and collecting the upper liquid. Drying at 40 deg.C (water content is less than or equal to 2%) to obtain primary refined tannin product with yield of 60.13% and total phenol content of 83.10%.
Taking 30g of HPD826 type macroporous adsorption resin, adding 95% ethanol, soaking for 24h to fully swell the macroporous adsorption resin, filtering, washing with a large amount of distilled water until the filtered solution has no obvious alcohol smell, and soaking with distilled water for storage.
30g of the treated HPD826 type resin is taken and loaded into a chromatographic column by a wet method, and 4.0BV of tannin solution of 2.00g/L prepared by the above primary refined tannin sample is added for adsorption at a flow rate of 1.5 BV/h. After the adsorption equilibrium, the column was washed with 2.0BV of distilled water and eluted with 1.6BV of 70% ethanol solution at a flow rate of 1.5 BV/h. Collecting eluate, concentrating at 50 deg.C with rotary evaporator, and drying at 40 deg.C (water content is less than or equal to 2%) to obtain refined tannin product with yield of 46.00%, total yield of refined tannin product of 27.66%, and total phenol content of refined tannin product of 99.97%.
Example 2
Adding petroleum ether into 10g of industrial myrica rubra tannin extract according to the mass-volume ratio of 1:2(g/mL), fully shaking up, degreasing for 30min under ultrasonic, filtering to remove the petroleum ether, and drying the myrica rubra tannin extract by blast at 40 ℃ (the water content is less than or equal to 2%). Adding 70mL of acetone into the degreased myrica rubra tannin extract, then slowly adding 30mL of distilled water, fully stirring for 30min, filtering by using a triangular funnel and collecting the filtrate. Removing acetone from the filtrate with a rotary evaporator at vacuum degree of 95-98 kPa and heating bath temperature of about 40 deg.C, collecting concentrated solution (mass concentration is controlled at about 30%), extracting with n-hexane for 1 time, collecting lower raffinate, extracting with chloroform for 1 time, and collecting upper liquid. Drying at 40 deg.C (water content is less than or equal to 2%) to obtain primary refined tannin product with yield of 60.13% and total phenol content of 83.10%.
Taking 30g of HPD826 type macroporous adsorption resin, adding 95% ethanol, soaking for 24h to fully swell the macroporous adsorption resin, filtering, washing with a large amount of distilled water until the filtered solution has no obvious alcohol smell, and soaking with distilled water for storage.
30g of the treated HPD826 type resin is taken and loaded into a chromatographic column by a wet method, and 4.0BV of tannin solution with the concentration of 3.00g/L prepared by the above primary refined tannin sample is added for adsorption at the flow rate of 1.5 BV/h. After the adsorption equilibrium, the column was washed with 2.0BV of distilled water and eluted with 1.6BV of 70% ethanol solution at a flow rate of 1.5 BV/h. Collecting eluate, concentrating at 50 deg.C with rotary evaporator, and drying at 40 deg.C (water content is less than or equal to 2%) to obtain refined tannin product with yield of 51.33%, total yield of refined tannin product of 30.86%, and total phenol content of refined tannin product of 96.73%.
Example 3
Adding petroleum ether into an industrial myrica rubra tannin extract 10 according to the mass-volume ratio of 1:2(g/mL), fully shaking up, degreasing for 30min under ultrasonic, filtering to remove the petroleum ether, and drying the myrica rubra tannin extract by blast at 40 ℃ (the water content is less than or equal to 2%). Adding 70mL of acetone into the degreased myrica rubra tannin extract, then slowly adding 30mL of distilled water, fully stirring for 30min, filtering by using a triangular funnel and collecting the filtrate. Removing acetone from the filtrate by using a rotary evaporator at the vacuum degree of 95-98 kPa and the heating bath temperature of about 40 ℃, collecting a concentrated solution (the mass concentration is controlled to be about 30%), extracting for 2 times by using n-hexane with the same volume, collecting the lower raffinate, extracting for 2 times by using chloroform with the same volume, and collecting the upper liquid. Drying at 40 deg.C (water content is less than or equal to 2%), to obtain refined tannin product with yield of 52.54% and total phenol content of 91.42%.
Taking 30g of HPD826 type macroporous adsorption resin, adding 95% ethanol, soaking for 24h to fully swell the macroporous adsorption resin, filtering, washing with a large amount of distilled water until the filtered solution has no obvious alcohol smell, and soaking with distilled water for storage.
30g of the above-mentioned treated HPD826 type resin was charged into a column by a wet method, and 4.0BV of a tannin solution (2.00 g/L) prepared from the above-mentioned preliminarily purified sample was added thereto and adsorbed at a flow rate of 1.5 BV/h. After the adsorption equilibrium, the column was washed with 2.0BV of distilled water and eluted with 1.6BV of 70% ethanol solution at a flow rate of 1.5 BV/h. Collecting eluate, concentrating at 50 deg.C with rotary evaporator, and drying at 40 deg.C (water content is less than or equal to 2%) to obtain refined tannin product with yield of 48.05%, total yield of the refined tannin product of 25.24%, and total phenol content of the refined tannin product of 100.00%.
Example 4
Adding petroleum ether into an industrial myrica rubra tannin extract 10 according to the mass-volume ratio of 1:2(g/mL), fully shaking up, degreasing for 30min under ultrasonic, filtering to remove the petroleum ether, and drying the myrica rubra tannin extract by blast at 40 ℃ (the water content is less than or equal to 2%). Adding 70mL of acetone into the degreased myrica rubra tannin extract, then slowly adding 30mL of distilled water, fully stirring for 30min, filtering by using a triangular funnel and collecting the filtrate. Removing acetone from the filtrate by using a rotary evaporator at the vacuum degree of 95-98 kPa and the heating bath temperature of about 40 ℃, collecting a concentrated solution (the mass concentration is controlled to be about 30%), extracting for 2 times by using n-hexane with the same volume, collecting the lower raffinate, extracting for 2 times by using chloroform with the same volume, and collecting the upper liquid. Drying at 40 deg.C (water content is less than or equal to 2%), to obtain refined tannin product with yield of 52.54% and total phenol content of 91.42%.
Taking 30g of HPD826 type macroporous adsorption resin, adding 95% ethanol, soaking for 24h to fully swell the macroporous adsorption resin, filtering, washing with a large amount of distilled water until the filtered solution has no obvious alcohol smell, and soaking with distilled water for storage.
30g of the above-mentioned treated HPD826 type resin was charged into a column by a wet method, and 6.0BV of a tannin solution (2.00 g/L) prepared from the above-mentioned preliminarily purified sample was added thereto and adsorbed at a flow rate of 1.5 BV/h. After the adsorption equilibrium, the column was washed with 2.0BV of distilled water and eluted with 1.6BV of 70% ethanol solution at a flow rate of 1.5 BV/h. Collecting eluate, concentrating at 50 deg.C with rotary evaporator, and drying at 40 deg.C (water content is less than or equal to 2%) to obtain refined tannin product with yield of 44.34%, total yield of 23.29% and total phenol content of 99.33%.
Example 5
Adding petroleum ether into an industrial myrica rubra tannin extract 10 according to the mass-volume ratio of 1:2(g/mL), fully shaking up, degreasing for 30min under ultrasonic, filtering to remove the petroleum ether, and drying the myrica rubra tannin extract by blast at 40 ℃ (the water content is less than or equal to 2%). Adding 70mL of acetone into the degreased myrica rubra tannin extract, then slowly adding 30mL of distilled water, fully stirring for 30min, filtering by using a triangular funnel and collecting the filtrate. Removing acetone from the filtrate by using a rotary evaporator at the vacuum degree of 95-98 kPa and the heating bath temperature of about 40 ℃, collecting a concentrated solution (the mass concentration is controlled to be about 30%), extracting for 2 times by using n-hexane with the same volume, collecting the lower raffinate, extracting for 2 times by using chloroform with the same volume, and collecting the upper liquid. Drying at 40 deg.C (water content is less than or equal to 2%), to obtain refined tannin product with yield of 52.54% and total phenol content of 91.42%.
Taking 30g of HPD826 type macroporous adsorption resin, adding 95% ethanol, soaking for 24h to fully swell the macroporous adsorption resin, filtering, washing with a large amount of distilled water until the filtered solution has no obvious alcohol smell, and soaking with distilled water for storage.
30g of the above-mentioned treated HPD826 type resin was charged into a column by a wet method, and 6.0BV of a tannin solution (3.00 g/L) prepared from the above-mentioned preliminarily purified sample was added thereto and adsorbed at a flow rate of 1.5 BV/h. After the adsorption equilibrium, the column was washed with 2.0BV of distilled water and eluted with 1.6BV of 70% ethanol solution at a flow rate of 1.5 BV/h. Collecting eluate, concentrating at 50 deg.C with rotary evaporator, and drying at 40 deg.C (water content is less than or equal to 2%) to obtain refined tannin product with yield of 41.12%, total yield of 21.60% and total phenol content of 97.14%.
The invention relates to a primary refined plant tannin product which is prepared by separating and purifying an organic solvent from industrial-grade myrica rubra tannin extract raw materials (first-grade myrica rubra tannin extract, the mass fraction of tannin is more than or equal to 67%, the total phenol content in the industrial myrica rubra tannin extract is 68.16% through the detection of a Prussian method, and the distance between the industrial-grade myrica rubra tannin extract and the technical standard of the industrial-grade myrica rubra tannin extract is very small), wherein the total phenol content of the primary refined plant tannin product reaches more than 83%. The preliminarily refined plant tannin product is subjected to further macroporous resin adsorption separation to finally obtain a high-purity plant tannin product, the total yield of the product is more than 20%, and the total phenol content can reach more than 96%.
Claims (10)
1. A method for preparing high-purity plant tannin by using industrial tannin extract is characterized by comprising the following steps:
(1) adding petroleum ether into industrial tannin extract, mixing well, degreasing under ultrasonic condition, separating, and collecting filtrate and residue respectively;
(2) drying the filter residue obtained in the step (1), adding acetone and distilled water into the dried filter residue, fully stirring, separating, and collecting filtrate;
(3) removing acetone from the filtrate collected in the step (2), collecting concentrated solution, extracting with n-hexane, collecting lower layer liquid, extracting with chloroform, collecting upper layer liquid, and drying to obtain primarily refined plant tannin solid;
(4) adding water into the primarily refined plant tannin solid obtained in the step (3) to prepare a primarily refined plant tannin solution;
(5) passing the primarily refined plant tannin solution prepared in the step (4) through a pretreated macroporous adsorption resin chromatographic column;
(6) after the adsorption balance in the step (5) is achieved, firstly, washing the chromatographic column with water, then, eluting with ethanol solution as an eluent, and collecting the eluent;
(7) and (4) concentrating the eluent collected in the step (6), and drying to obtain the plant tannin product.
2. The method of claim 1, wherein: the industrial-grade tannin extract in the step (1) is prepared by taking myrica rubra barks as raw materials, and the mass fraction of tannin is not less than 67%.
3. The method of claim 1, wherein: the boiling range of the petroleum ether used in the step (1) is 60-90 ℃, and the solid-to-liquid ratio of the industrial-grade tannin extract to the petroleum ether is 1:1-1:4(g/mL), and the preferred solid-to-liquid ratio is 1:2 (g/mL).
4. The method of claim 1, wherein: the degreasing time in the step (1) is 0.5-1.0h, and then filtration or centrifugal separation is carried out to respectively obtain filtrate and filter residue; the ultrasonic power range 240-.
5. The method of claim 1, wherein: and (2) drying the filter residue obtained in the step (1) until the water content is less than or equal to 2%, wherein the volume ratio of acetone to distilled water is 1:9-9:1, the total solid-liquid ratio is 1:5-1:15(g: mL), preferably the volume ratio of acetone to distilled water is 7:3, the total solid-liquid ratio is 1:10(g: mL), fully stirring for 0.5-1.0h, filtering or centrifugally separating, and collecting filtrate.
6. The method of claim 1, wherein: and (3) removing acetone from the filtrate obtained in the step (2) by using a rotary evaporator at the vacuum degree of 95-98 kPa and the heating bath temperature of 38-42 ℃, collecting a concentrated solution with the mass concentration controlled at 28-32%, extracting for at least 1 time by using n-hexane with the same volume, collecting a lower layer of liquid, extracting for at least 1 time by using chloroform with the same volume, collecting an upper layer of liquid, and then drying in vacuum at the temperature of 30-60 ℃, preferably 40 ℃ until the water content is less than or equal to 2%, thus obtaining a reddish black solid, namely the primarily refined plant tannin solid.
7. The method of claim 1, wherein: in the step (4), the preliminarily refined plant tannin solid sample obtained in the step (3) is added with water to prepare a solution with the mass concentration of 1.0g/L-5.0g/L, preferably 1.5g/L-3.0 g/L.
8. The method of claim 1, wherein: step (5) putting the pretreated macroporous resin into a chromatographic column by a wet method, adding the preliminarily refined plant tannin solution prepared in the step (4) of 1.0BV-6.0BV, and adsorbing at the flow rate of 1.0BV/h-4.0 BV/h; preferably, 4.0BV of the primarily refined plant tannin solution prepared in the step (4) is added, and the adsorption is carried out at the flow rate of 1.5 BV/h;
after the adsorption balance is achieved in the step (6), washing the chromatographic column by distilled water with the volume of 1.0BV-4.0BV, then eluting by using ethanol solution with the volume fraction of 1.0BV-3.0BV being 40% -90% as an eluent with the flow rate of 1.0BV-3.0BV/h, and collecting eluent; preferably 2.0BV volume of distilled water is used to wash the column, preferably 1.6BV volume fraction 70% ethanol solution is used as eluent, and the flow rate is preferably 1.5 BV/h.
9. The method of claim 1, wherein: and (7) concentrating the eluent obtained in the step (6) at 30-60 ℃ by using vacuum drying equipment, preferably at 50 ℃, and then drying at 30-60 ℃, preferably at 40 ℃ until the water content is less than or equal to 2% to obtain a high-purity plant tannin solid sample.
10. A high purity vegetable tannin produced by the method of any one of claims 1 to 9.
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